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基因枪法转Bt/GNA基因超甜玉米植株的获得及抗虫性鉴定 总被引:1,自引:0,他引:1
以超甜玉米自交系S1幼胚诱导的愈伤组织为受体材料,利用基因枪法共转化Bt和GNA基因,获得了128株再生植株,移栽成活31株,其中26株可育.经PCR检测外源基因整合进玉米基因组中,含Bt基因11株,其中有5株能扩增出GNA基因.Southern杂交证明,目的基因已经整合在玉米基因组中.对9株转基因T1代植株叶片室内接虫抗性鉴定,结果显示,5株转基因植株叶片饲喂6 d后明显抑制玉米螟幼虫的发育,幼虫体重增加较少,幼虫死亡率高,表现较强的抗性. 相似文献
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采用本研究室建立的根癌农杆菌介导的高效遗传转化体系,转化苎麻优良品种芦竹青,获得了转Bt基因苎麻候选植株。通过PCR和Southern杂交等分子检测,证明Bt基因已经整合到部分候选植株基因组中。选取PCR和Southern杂交均为阳性的部分株系(T0)种植在大田,对这些植株进行室内抗虫鉴定并考察其主要农艺性状和品质性状,次年对T1代植株进行PCR和Southern杂交检测。结果表明, 与对照相比,T0代转Bt基因苎麻植株的抗虫性均强于对照,部分株系的抗虫性显著强于对照植株;且基本保持了亲本的优良性状;T1代植株中也含有Bt基因,表明Bt基因能稳定遗传,且T1代植株在大田的抗虫性明显强于非转基因植株。 相似文献
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转Bt基因抗虫棉育种策略与效果 总被引:17,自引:3,他引:17
应用改进的常规育种技术培育转Bt基因抗虫棉,可大大提高对鳞翅目害虫的抗性,这是棉花品种改良的新趋势。育种实践表明,根据转Bt基因种质系供体的特点和遗传缺陷,选择综合性状优良、前期生长势强、株形稍高大松散,铃重较大、生育期相对较短的材料作亲本较为适宜;在杂交技术的应用上,宜采用复合杂交、基因渐渗和有利基因累加技术;杂种后代的处理视杂交方式而定。本文对育种程序中抗虫性鉴定技术进行了讨论。 相似文献
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转Bt基因抗虫棉抗虫性遗传研究 总被引:13,自引:4,他引:13
以转 Bt基因抗虫棉 R55为材料 ,利用ELISA( Enzyme-linked Immunosorbent Assay)检测方法 ,通过对不同亲本与抗虫亲本 R55杂交F1~ F5、BC1、BC2 世代材料 Bt晶体杀虫蛋白的定性、定量测定 ,结合大田自然感虫条件下的抗棉铃虫鉴定 ,研究了转 Bt基因抗虫棉 Bt基因的遗传规律。结果表明 :F1均表现阳性 ,F2 阳、阴性株符合 3∶ 1的分离比例 ,回交 BC1阳、阴性株呈现 1∶ 1的分离比例 ,说明 Bt基因的遗传基本符合显性主基因遗传规律。但 Bt基因的遗传又有其特殊性 ,表现在亲本的遗传背景对 Bt基因的表达有着较大影响 ,不同亲本与抗虫亲本杂交 F1代 Bt晶体蛋白的表达量存在较大差异 ;杂交方式对 Bt基因的表达亦有一定影响 ;不加选择的连续回交 ,有可能使 Bt基因“丢失”。未发现 Bt基因的表达随世代的递增、农艺和经济性状的改进而降低的趋势 ,Bt基因可以稳定遗传 相似文献
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转Bt基因棉及其抗虫性研究与利用进展 总被引:26,自引:2,他引:24
本文综述了转Bt基因棉的培育及应用,对鳞翅目害虫的抗性及对非鳞翅目害虫和天敌的影响,以及害虫对转Bt基因棉抗性的预防和治理等方面的研究进展。并针对我国的具体情况,讨论了害虫对转Bt基因棉抗性的预防策略及转基因棉IPM体系的关键技术组成。 相似文献
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分子标记辅助选择聚合棉花Rf1育性恢复基因和抗虫Bt基因 总被引:19,自引:0,他引:19
胞质雄性不育恢复系0-613-2R与转Bt基因抗虫棉R019(轮回亲本)杂交、回交产生BC2群体。利用CMS恢复基因Rfl紧密连锁的3个SSR标记和Bt基因的PCR标记开展分子标记辅助选择培育聚合有Rfl和Bt的转基因抗虫棉恢复系。在分析的59个BC2单株中55株存在恢复基因标记,54株存在Bt基因;综合标记分析结果,共获得54个同时具Rfl与Bt基因的聚合单株;这些聚合单株自交后,通过标记辅助选择,获得10株含Bt基因且Rfl纯合的单株。为棉花优良恢复系的快速培育提供了重要基础。 相似文献
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反式烯脂酰-CoA还原酶(trans-2, 3-enoyl-CoA reductase, ECR)是催化超长链脂肪酸(VLCFAs)合成的脂肪酰-CoA延长酶之一。根据已报道拟南芥等的ECR基因设计引物,采用RACE (rapid amplification of cDNA ends)方法从甘蓝型油菜中克隆ECR的全长cDNA序列和对应的基因组序列,命名为BnECR (GenBank登录号分别为FJ899705和FJ899706)。序列分析结果显示,BnECR的全长cDNA序列为1 328 bp,对应的基因组序列为2 093 bp,由4个外显子组成,在ORF的上、下游分别有一个163 bp的5′UTR和一个232 bp的3′UTR。根据编码区预测BnECR前体蛋白为一个310个氨基酸残基的多肽链,包含ECR蛋白的重要功能位点K144、R145及一个NAD(P)H结合基序G225SGGYQIPR/HG234。NCBI Blastn、氨基酸序列多重比对及保守域分析表明, 该基因与拟南芥AtECR基因的同源性最高,是对应拟南芥AtECR的垂直同源基因。RT-PCR分析表明,BnECR基因在甘蓝型油菜根、茎、叶、花及角果中均有表达,其中在茎中的表达量最高。BnECR在高芥酸材料种子发育中后期的表达量显著高于低芥酸种子,表明BnECR可能参与甘蓝型油菜芥酸的合成。将BnECR克隆到酿酒酵母的穿梭表达载体中,分别转化野生型酵母By4743和突变体菌株YDL015c,添加半乳糖诱导表达。气相色谱分析表明,BnECR在酿酒酵母中有效表达,转化菌株中的芥酸(C22:1)占总脂肪酸含量的1.34%,比对照增加了52%;对突变体的转化结果表明芥酸含量恢复到野生型水平。 相似文献
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Studies were undertaken to induce early flowering mutants by ethyl methanesulphonate (EMS) treatments of Brassica napus seeds. EMS treatments for 12 h of a highly inbred B. napus line TBS had adverse effects on M1 plant development and fertility only when concentrations were greater than 1%. However, an EMS concentration of 1.5% did not reduce M1 plant fertility to an extent which significantly reduced production of M2 seeds. Genetic changes induced by EMS treatment and affecting flowering time were of three main types: (1) Changes within a polygenic system reflected by increased variation in flowering time among M2 families. As the increase in variation was due primarily to a higher frequency of later flowering plants, these polygenic changes would be of little value in developing better-adapted cultivars. (2) Induction of a recessive mutation at a major gene locus which caused M3 plants homozygous for the mutant gene to flower at least 20 days earlier than the parental line TB8. (3) Induction of a dominant mutation at a major gene locus which affects flowering time by causing a substantial reduction in vernalization requirement. M2 plants carrying the mutant gene flowered as early as 59 days before the parental line. These major gene mutations could be rapidly exploited in the development of agronomically superior cultivars for short-season, lower rainfall environments in Western Australia. 相似文献
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转Bt基因棉杀虫蛋白含量时空分布及对棉铃虫产生抗性的影响 总被引:17,自引:9,他引:17
采用ELISA法(酶链免疫法)、室内初孵棉铃虫生测法和田间棉铃虫为害调查方法,研究和分析了中国构建的 Bt(Bacillus thuringiensis)基因抗虫棉品系GK3和美国构建的Bt基因棉品系新棉33B的不同生育期以及花铃期不同器官的杀虫蛋白含量、校正死亡率和田间受害率变化趋势以及它们之间的关系。结果表明,转Bt基因棉Bt杀虫蛋白含量在棉花生育过程中呈时空动态变化,在时间分布上,各生育期顶尖平展叶表现为:初花期>蕾期、花铃期>苗期>吐絮期;在花铃期,各器官表现为:功能叶、茎尖>小蕾、幼铃>花蕊、花瓣、苞叶、老叶。室内生测幼虫校正死亡率与棉株Bt杀虫蛋白含量高度一致;田间表现与Bt杀虫蛋白含量有一定的差异,除主要受Bt杀虫蛋白影响外,棉铃虫取食选择性,以及残存高龄幼虫为害和棉株各部位营养结构等也影响其抗虫性,造成后期棉铃虫主要为害花、蕾和幼铃,两材料抗虫性表现较为一致。 相似文献
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甘蓝型油菜PIN家族基因的鉴定与生物信息学分析 总被引:1,自引:0,他引:1
PIN家族基因是一类调控植物生长素极性运输的重要载体元件, PIN基因编码生长素输出蛋白, 介导生长素在植物体的运输, 然而在基因组较复杂的甘蓝型油菜中缺乏系统研究。本研究运用生物信息学方法在甘蓝型油菜全基因组数据库筛选甘蓝型油菜PIN家族基因, 对鉴定出的29个BnPINs基因开展拷贝数变异、分子特征、跨膜结构域、保守基序、染色体定位、系统进化树构建、PIN蛋白二级结构及三级结构预测等研究, 结合高通量转录组测序进行低氮胁迫下的转录水平分析。结果表明, 甘蓝型油菜PIN家族基因拷贝数明显多于拟南芥、甘蓝和白菜所具有的PIN家族基因数量; BnPINs蛋白多属于由碱性氨基酸组成的稳定蛋白, 含有保守的N末端结构域, 二级结构与拟南芥PIN蛋白相似; 系统进化选择能力分析表明, BnPINs基因与甘蓝和白菜PIN家族基因进化关系相近。转录组测序表明, BnPIN1s、BnPIN2s、BnPIN3s基因主要在甘蓝型油菜根部表达且受长期低氮(72 h)诱导, BnPIN6s和BnPIN8s基因主要在地上部表达, 低氮会抑制BnPIN6s表达。本研究结果为进一步研究甘蓝型油菜PIN家族基因生物学功能尤其是在响应低氮胁迫中的功能奠定基础, 为已知大量数据的其他物种家族基因生物信息学研究提供参考。 相似文献
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Ten quite different European winter rapeseed cultivars were grown in a 3-year field trial at one location. The traits grain yield and biomass were measured based on single plant measurements (first year) or on plot basis (second and third year). Individual harvest indices were calculated.
Besides a general discussion of the experimental results for this collection of winter rapeseed cultivars the main aim of the present investigations was an examination whether these experimental results are in agreement with the theoretical conclusions on harvest indices which have been published in this journal in three preceding publications.
Hence the investigations of this paper are divided into the three chapters: 1) Calculation of mean harvest indices, 2) Variability of harvest indices and 3) Correlations among harvest index, grain yield and biomass. In all three chapters the agreement between the empirically obtained and the theoretically expected values is quite good. 相似文献
Besides a general discussion of the experimental results for this collection of winter rapeseed cultivars the main aim of the present investigations was an examination whether these experimental results are in agreement with the theoretical conclusions on harvest indices which have been published in this journal in three preceding publications.
Hence the investigations of this paper are divided into the three chapters: 1) Calculation of mean harvest indices, 2) Variability of harvest indices and 3) Correlations among harvest index, grain yield and biomass. In all three chapters the agreement between the empirically obtained and the theoretically expected values is quite good. 相似文献
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此文首先回顾了国内外抗虫转基因玉米研究发展历程,归纳总结了转Bt毒蛋白基因玉米的常用转化方法和转基因玉米转化体的鉴定方法,对转Bt毒蛋白基因玉米后续实验的抗虫性分析鉴定及遗传稳定性评价与利用也进行了介绍。进而探讨了Bt毒蛋白基因在玉米遗传转化上善待解决的问题以及未来的发展方向。认为应加大力度对瓶颈技术进行深入研究,以期转Bt毒蛋白基因玉米能够获得更好的发展。 相似文献
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Periods of drought frequently affect the development of winter rape at the shooting stage in spring. A growth chamber study was conducted to determine the effects of 13 days of strong water deficit at the shooting stage followed by rewatering on oilseed rape (Brassica napus). The osmolality and the efficiency of photosystem II in leaves were measured by means of a vapour pressure osmometer and a chlorophyll fluorometer, respectively. The activities of sucrose cleaving enzymes in leaves were determined through invertase as well as sucrose synthase tests. A higher osmotic pressure was observed 2 days after water deprivation. Extracellular and vacuolar invertases as well as chlorophyll fluorescence were decreased with delay. Rewatering led to a gradual decrease in the osmotic pressure and a delayed increase in the enzyme activities as well as the photosynthetic efficiency. A linear regression model could predict osmolality in well‐watered plants based on the extracellular enzyme activity and growth stage. The ontogenetic and stress‐related pattern of extracellular invertase activity could indicate a role of this enzyme in balancing source‐sink relations. The onset of flowering was not influenced by drought, but the period of this stage was prolonged. Though, the ontogenetic stages of stressed and unstressed plants were the same at the time of harvest, the development of seed biomass was significantly depressed under stress. 相似文献